番茄中与致病密切相关miRNA的挖掘及特性分析
收稿日期: 2013-08-07
修回日期: 2013-09-24
网络出版日期: 2013-12-20
基金资助
国家自然科学基金项目(编号：30972001, 31272167) 资助
Mining and characterization of miRNAs closely associated with the pathogenicity in tomato
Received date: 2013-08-07
Revised date: 2013-09-24
Online published: 2013-12-20
MicroRNAs(miRNAs)是一类在真核生物体内普遍存在的、长度约22nt的内源性非编码小分子RNA, 它通过与靶mRNA的结合参与多种基因的表达调控。番茄作为一种重要的模式植物, 其miRNA的研究近年来也取得了较大的进展。文章通过搜集已报道的文献和miRBase, 找到番茄中34个miRNA的表达与致病相关, 采用生物信息学方法预测它们的靶基因, 利用Cytoscope软件构建miRNA及其靶基因的调控网络。从中筛选出13个与致病密切相关的miRNA, 根据各miRNA之间关联性的大小及其与致病相关靶基因的多少, 进一步选出miR169、miR482、miR5300、miR6024、miR6026和miR6027, 并对其进行了靶基因功能分析、启动子分析及实时定量PCR验证, 为全面深入研究miRNA的作用机制奠定了基础。
孙广鑫, 栾雨时, 崔娟娟 . 番茄中与致病密切相关miRNA的挖掘及特性分析[J]. 遗传, 2014 , 36(1) : 69 -76 . DOI: 10.3724/SP.J.1005.2014.00069
MicroRNAs (miRNAs) are a class of small endogenously RNA with an approximate length of 22nt which are known to be ubiquitous in eukaryotes. More importantly, miRNAs are key regulators of gene expression in eukaryotic cells through the degradation of target mRNA. The investigation on miRNAs in tomato that is an important model plant has made a great progress in recent years. Herein, by collecting the reported literature and miRBase, we found that 34 miRNAs in tomato are closely associated with pathogenicity. Subsequently, we predicted their target genes through bioinformatics approaches and built a disease-related regulatory network of miRNA and its target genes using Cytoscope program. This has led us to identify 13 miRNAs that are closely associated with pathogenicity in tomato from which we selected miR169, miR482, miR5300, miR6024, miR6026 and miR6027 for further analysis based on the association between miRNAs and the number of target genes. Lastly, we performed the analysis of target gene, promoter and real time quantitative PCR verification for these 6 miRNAs. Our study may pave the way for future in-depth analysis of biological action of miRNAs.
Key words： miRNA; tomato; bioinformatics; target gene; promoter
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